Identification and Removal of Nitroxide Spin Label Contaminant: Impact on PRE Studies of α-helical Membrane Proteins in Detergent

Overview

Journal Protein Sci

Specialty Biochemistry

Date 2012 Mar 6

PMID 22389096

Citations 3

Authors

Brett M Kroncke

Linda Columbus

Affiliations

Soon will be listed here.

Abstract

NMR paramagnetic relaxation enhancement (PRE) provides long-range distance constraints (~15-25 Å) that can be critical to determining overall protein topology, especially where long-range NOE information is unavailable such as in the case of larger proteins that require deuteration. However, several challenges currently limit the use of NMR PRE for α-helical membrane proteins. One challenge is the nonspecific association of the nitroxide spin label to the protein-detergent complex that can result in spurious PRE derived distance restraints. The effect of the nitroxide spin label contaminant is evaluated and quantified and a robust method for the removal of the contaminant is provided to advance the application of PRE restraints to membrane protein NMR structure determination.

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